Suppression of vascular endothelial cell inflammation Both adaptive and innate immunity tightly regulate atherosclerosis. Vascular endothelial cells (ECs) are targets of proinflammatory cytokines, such as interleukin-1b (IL-1b) and tumor necrosis factor-a (TNF-a). However, the important question of whether ECs play a critical role in initiating vascular inflammation remains poorly defined. In particular, the issue of whether ECs in atherogenic conditions generate excess IL-1b via the newly-defined inflammasome remains unknown. Therefore, the goal of this proposal is to examine a novel hypothesis that EC inflammation with the generation of IL-1b followed by EC apoptosis is accelerated in the pathogenesis of atherosclerosis. The corollary hypothesis is that factors and cells in suppressing EC generation of excess IL- 1b and EC apoptosis play inhibitory roles in atherogenesis. Preliminary data from our lab and reports from others' labs clearly support this hypothesis: (i) Microbial infections play a role in the pathogenesis of atherosclerosis. In response to the stimulation of bacterial lipopolysaccharide (LPS), mouse aortic ECs (MAECs) from atherogenic apolipoprotein E-deficient ApoE-/- mice express higher levels of pro-IL-1b than MAECs from wild-type control mice. The upregulation of pro-IL-1b in MAECs from ApoE-/- mice is increased in the presence of phosphatidylinositol 3-kinase (PI-3K) inhibitors Wortmannin and Ly294002, suggesting that PI3K-Akt survival pathway suppresses IL-1b-generating EC inflammation; (ii) We have identified a novel Bcl-xL interacting, anti-apoptotic protein TCTP. TCTP is an important regulator and anti-apoptotic protein along PI3K- Akt survival pathway. In addition, TCTP is also down-regulated in response to LPS stimulation in ECs, suggesting that anti-TCTP may inhibit EC inflammation and apoptosis as a PI-3K-Akt pathway-signaled protective mechanism of ECs. In addition, we found that co-culture of MAECs from ApoE-/- mice with CD4+CD25highFoxp+ regulatory T cells (Tregs) results in decreased pro-IL-1b expression in response to LPS, suggesting that immunosuppressive Tregs suppress EC inflammation; (iii) IL-1b deficiency inhibits EC inflammation and atherogenesis. Therefore, the elucidation of the suppressive mechanisms of IL-1b-generating EC inflammation and EC apoptosis is clinically significant, as it may lead to new therapeutics for treating atherosclerosis and vascular inflammation. This goal will be pursued through the execution of the following specific aims: (1) To determine whether IL-1b-generating EC inflammation is accelerated in atherogenic ApoE-/- mice in comparison to that in wild-type control mice; (2) To determine whether TCTP and PI3K-Akt intracellular signaling pathway inhibits IL-1b-generating EC inflammation and EC apoptosis in ApoE-/- mice; and (3) To determine whether extracellular suppression of IL-1b-generating EC inflammation and EC apoptosis by Tregs attenuates the atherogenesis in ApoE-/- mice.